C型钠肽和生长分化因子9预处理对绵羊体外成熟卵母细胞质量和发育能力的影响  

Effects of pre-treatment with C-type natriuretic peptide and growth differentiation factor 9 on the quality and developmental potential of in vitro matured sheep oocytes

作　　者：崔健 唐雅雯 王兆琛 张家新[2] 仇春娟 包梅荣 安磊[1] 田见晖[1] CUI Jian;TANG Yawen;WANG Zhaochen;ZHANG Jiaxin;QIU Chunjun;BAO Meirong;AN Lei;TIAN Jianhui(College of Animal Science and Technology,China Agricultural University,Beijing 100193,China;College of Animal Science and Technology,Inner Mongolia Agricultural University,Hohhot O10010,China;Inner Mongolia Sino Sheep Technology Co.,Ltd.,Ulanqab,Inner Mongolia 011800,China)

机构地区：[1]中国农业大学动物科学技术学院,北京100193 [2]内蒙古农业大学动物科学技术学院,内蒙古呼和浩特010010 [3]内蒙古赛诺种羊科技有限公司,内蒙古乌兰察布011800

出　　处：《中国兽医学报》2023年第3期610-621,629,共13页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31972573,31672426);国家重点研发计划资助项目(2021YFD1300901,2017YFD0501901,2017YFD0501905);农业产业技术体系北京市创新团队资助项目。

摘　　要：为探讨生长分化因子9(growth differentiation factor 9,GDF9)和骨形态发生蛋白15(bone morphogenetic protein 15,BMP15)在协同C型钠肽(C-type natriuretic peptide,CNP)调节卵母细胞减数分裂进程中的作用,提高卵母细胞体外成熟的质量,试验基于CNP预处理的“两步法”体外成熟(in vitro maturation,IVM)体系,研究了预处理阶段添加GDF9和BMP15在调节绵羊卵母细胞减数分裂进程和胞质成熟事件中的作用,并探讨CNP和GDF9联合预处理后对卵母细胞体外成熟质量及发育能力的影响。结果显示,GDF9可显著提高卵丘细胞中CNP受体NPR2基因的表达,从而增加CNP阻滞绵羊卵母细胞减数分裂的效率,而BMP15无此作用。预处理阶段联合添加CNP和GDF9能改善卵丘细胞功能,并且在CNP的作用基础上进一步增加卵母细胞线粒体的活性和数量、GSH含量,并降低ROS水平,从而提高卵母细胞体外成熟后的质量以及体外受精后的发育效率,卵裂率和囊胚率得到显著增加。研究结果不仅表明GDF9能强化CNP在卵母细胞的细胞核与细胞质同步成熟中的作用,也为优化绵羊的体外胚胎生产体系提供了新的方法。The aim of this study is to determine the role of growth differentiation factor 9(GDF9)and bone morphogenetic protein 15(BMP15)in coordinating C-type natriuretic peptide(CNP)-mediated oocyte meiotic progression,and to improve the quality of in vitro matured oocytes.Using CNP pre-treatment-based“two-step”in vitro maturation(IVM)system,we investigated the effect of pre-treatment with CNP,in combination with GDF9 and/or BMP15,on meiotic progression and cytoplasmic maturation of sheep oocytes.In addition,we tested the effects of pre-treatment with CNP and GDF9 on the quality and developmental potential of in vitro matured sheep oocytes.The results showed that GDF9,but not BMP15,significantly increased the efficiency of CNP pre-treatment in blocking meiotic resumption of sheep oocytes.Combined pre-treatment with CNP and GDF9,not only improved the status of cumulus cells,but also increased mitochondrial mass and activity,GSH content,and reduced ROS levels in oocytes,thereby improving the quality of in vitro maturated sheep oocytes and their developmental efficiency following in vitro fertilization,cleavage and blastocyst rates were significantly increased.Collectively,our study not only identifies that GDF9can enhance the efficacy of CNP in the synchronizing nuclear and cytoplasmic maturation of sheep oocytes,but also presenting a novel method for improving in vitro embryo production system of ruminants.

关 键 词：绵羊 卵母细胞 “两步法”体外成熟 C型钠肽 生长分化因子9 

分 类 号：S816[农业科学—饲料科学]